anyone using TRIzol from invitrogen? - (Jun/20/2005 )

hi,anyone using TRizol for RNA exaction? it is written in the protocol that there will be three layers after phase separation: a lower red phenol-chloroform phase, an interphase and a colorless upper aqueous phase. but every time i used TRIzol, i only have two layers: the lower red phenol-chloroform phase and the colorless upper aqueous phase. there were some white stuff btw two layers and i don't know what they are. do you guys ever had the same problem? does that mean i have to add more TRIzol reagent?

-hsm142-

The white stuff between the upper, clear layer and the lower red phase? yah, that is your third phase. It looks like that for me too. That is where the protein is. Avoid it when you are extracting the upper layer.

-pBluescript-

hithe so called third layer is dependant of the amount of cell/tissue you started from. the more sample you have at beginning the more important this phase will be.